Lipid Droplet Assay in Huh7 Cells

Lipid droplet assay was performed according to a previous method using a BSA-conjugated oleic acid system in Huh7 cells as described before (Yen et al., 2018 (link)). Briefly, cells seeded in μClear^® 96-well plates (Greiner Bio-ONE, Frickenhausen, Germany) were treated with oleic acid and the tested samples or DMSO for 18 h. Cells were stained with 2 μg/ml Hoechst 33342 and 1 μg/ml BODIPY^® 493/503 and fixed in paraformaldehyde. A High-content imaging (HCS) instrument was used to take and analyze images of the nuclei and lipid droplets (ImageXpress Micro System, Molecular Devices, Sunnyvale, CA, United States). The diameter settings were 8–25 μm for the nuclei and 0.5–2 μm for the lipid droplets.

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Korinek M., Handoussa H., Tsai Y.H., Chen Y.Y., Chen M.H., Chiou Z.W., Fang Y., Chang F.R., Yen C.H., Hsieh C.F., Chen B.H., El-Shazly M, & Hwang T.L. (2021). Anti-Inflammatory and Antimicrobial Volatile Oils: Fennel and Cumin Inhibit Neutrophilic Inflammation via Regulating Calcium and MAPKs. Frontiers in Pharmacology, 12, 674095.

Publication 2021

μClear® 96-well plates BODIPY® 493/503 ImageXpress Micro System

Assay Bodipy 493 503 Cells Devices Dmso Hoechst 33342 Lipid droplets Nuclei Oleic acid Paraformaldehyde

Corresponding Organization : Ming Chi University of Technology

Other organizations : Kaohsiung Medical University, National Sun Yat-sen University

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Variable analysis

independent variables

Oleic acid
Tested samples

dependent variables

Lipid droplet formation

control variables

Hoechst 33342 staining
BODIPY® 493/503 staining
Paraformaldehyde fixation
Nucleus diameter (8-25 μm)
Lipid droplet diameter (0.5-2 μm)

controls

Positive control: Oleic acid
Negative control: DMSO

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